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Generation of Stable and Unmarked Conditional Mutants in Pseudomonas aeruginosa

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Lipopolysaccharide Transport

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2548))

Abstract

The functional and physiological characterization of bacterial genes required for growth and/or cell survival is limited by the inability to generate deletion mutants lacking the specific gene of interest. This limitation can be circumvented by generating conditional mutants in which the loss of the endogenous copy of the gene is compensated by the introduction of the wild-type allele under the control of an inducible promoter, which allows for tightly regulated expression of the gene of interest. Besides the confirmation and/or functional investigation of essential genes, conditional mutants can also be useful to investigate the effect of finely controlled expression of nonessential genes. In this chapter, we describe a method that can be used to generate stable and unmarked conditional mutants in Pseudomonas aeruginosa.

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Acknowledgments

The research on essential genes in our laboratory was supported by the Pasteur Institute-Cenci Bolognetti Foundation, PRIN 2020 (grant protocol 20208LLXEJ), the Excellence Departments grant from the Italian Ministry of Education, University and Research (MIUR, Italy) (Art. 1, commi 314-337 Legge 232/2016), and the Regione Lazio (“Gruppi di Ricerca 2020,” POR A0375E0026).

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Authors and Affiliations

  1. Department of Science, University Roma Tre, Rome, Italy

    Alessandra Lo Sciuto, Maria Concetta Spinnato & Francesco Imperi

  2. Department of Biology and Biotechnology “Charles Darwin”, Sapienza University of Rome, Rome, Italy

    Martina Pasqua

  3. IRCCS Fondazione Santa Lucia, Rome, Italy

    Francesco Imperi

Authors
  1. Alessandra Lo Sciuto
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  2. Maria Concetta Spinnato
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  3. Martina Pasqua
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  4. Francesco Imperi
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Corresponding author

Correspondence to Francesco Imperi .

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Editors and Affiliations

  1. Department of Pharmacological and Biomolecular Sciences, Università degli Studi di Milano, Milan, Italy

    Paola Sperandeo

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Lo Sciuto, A., Spinnato, M.C., Pasqua, M., Imperi, F. (2022). Generation of Stable and Unmarked Conditional Mutants in Pseudomonas aeruginosa. In: Sperandeo, P. (eds) Lipopolysaccharide Transport. Methods in Molecular Biology, vol 2548. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2581-1_2

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  • DOI: https://doi.org/10.1007/978-1-0716-2581-1_2

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2580-4

  • Online ISBN: 978-1-0716-2581-1

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